Modified recombinant collagenase, compositions comprising the same and uses thereof in dental related procedures

Abstract

Provided herein are modified forms of recombinant collagenase polypeptide having one or more amino acid substitutions and/or deletions compared to a wild type recombinant collagenase protein. Further provided are nucleic acid molecules encoding the modified recombinant collagenase polypeptide, compositions including the same and uses thereof in various dental related procedures.

1 .- 29 . (canceled) 
     
     
         30 . A modified recombinant collagenase polypeptide, said modified recombinant collagenase polypeptide comprising one or more amino acid replacements relative to a corresponding wild type collagenase amino acid sequence; and wherein the thermal stability of the modified recombinant collagenase polypeptide is higher than the thermal stability of a corresponding wild type (WT) polypeptide. 
     
     
         31 . The modified recombinant collagenase polypeptide according to  claim 30 , comprising an N-terminal truncation, wherein the N-terminal truncation comprises a truncation of at least 15 amino acids relative to the corresponding wild type collagenase amino acid sequence. 
     
     
         32 . The modified recombinant collagenase polypeptide according to  claim 30 , comprising an amino acid sequence as denoted by any one of SEQ ID NO: 3, 5, 7, 13, 15 or 17. 
     
     
         33 . The modified recombinant collagenase polypeptide according to  claim 30 , comprising an amino acid sequence as denoted by SEQ ID NO: 3 or SEQ ID NO: 13. 
     
     
         34 . The modified recombinant collagenase polypeptide according to  claim 30 , wherein the polypeptide further comprises a Tag sequence at the N-terminus and/or the C-terminus thereof. 
     
     
         35 . The modified recombinant collagenase polypeptide according to  claim 30 , wherein the thermal stability of the modified recombinant collagenase polypeptide is higher than the thermal stability of a recombinant polypeptide comprising an N-terminal truncation. 
     
     
         36 . A composition comprising the modified recombinant collagenase polypeptide according to  claim 30 . 
     
     
         37 . A method of treating a dental related condition in a subject in need thereof, the method comprising administering to the subject in need thereof an effective amount of the modified recombinant collagenase polypeptide according to  claim 30 , or a composition comprising the same. 
     
     
         38 . The method according to  claim 37 , wherein the dental related procedures is selected from tooth extraction, orthodontic procedure and implant removal. 
     
     
         39 . The method according to  claim 37 , wherein the modified recombinant collagenase polypeptide or the composition is administered prior to, during or after the dental related procedure. 
     
     
         40 . The method according to  claim 37 , wherein the modified recombinant collagenase polypeptide or the composition comprising the same is administered locally. 
     
     
         41 . The method according to  claim 40 , wherein the administration is by local injection. 
     
     
         42 . The method according to  claim 37 , wherein administration thereof reduces extraction force during tooth extraction, wherein the required extraction force is reduced compared to administration of a WT collagenase (having an amino acid sequence as denoted by SEQ ID NO: 10), a truncated recombinant collagenase (having an amino acid sequence as denoted by SEQ ID NO: 1), or to administration of a carrier. 
     
     
         43 . The method according to  claim 42 , wherein the modified recombinant collagenase polypeptide or the composition comprising the same are administered locally to a periodontal ligament (PDL) region prior to the tooth extraction. 
     
     
         44 . A nucleic acid molecule encoding the modified recombinant collagenase of  claim 30 . 
     
     
         45 . The nucleic acid molecule according to  claim 44 , comprising a nucleotide sequence as denoted by any one of: SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 14, SEQ ID NO: 16, or SEQ ID NO: 18. 
     
     
         46 . A vector comprising the nucleic acid molecule of  claim 44 . 
     
     
         47 . A host cell comprising the nucleic acid molecule according to  claim 44 . 
     
     
         48 . A host cell transformed or transfected with the vector according to  claim 46 . 
     
     
         49 . A host cell comprising the modified recombinant collagenase polypeptide according to  claim 30 .